Biofortification of lines of pearl millet (Pennisetum glaucum (L.) R.Br.) with increased iron (Fe) and zinc (Zn) will have great impact because pearl millet is an indispensable component of food and nutritional security of inhabitants of arid and semi-arid regions. The aim of the present study was to assess the stability of Fe and Zn content in recombinant inbred lines (RILs) developed for grain Fe and Zn content, and to use these lines in developing micronutrient-rich pearl millet hybrids. A mapping population consisting of 210 RILs along, with parents and checks, was assessed in three consecutive years (2014–16) under rainfed conditions at the same experimental location in an alpha design with two repetitions. Significant differences were observed in genotype, environment and genotype × environment interaction mean squares for all variables, particularly grain micronutrients. The first two principal components of an interaction principal component analysis cumulatively explained 100% of the total variation; respective contributions of the first and second components were 64.0% and 36.0% for Fe, and 58.1% and 41.9% for Zn. A positive and moderately high correlation (0.696**) between Fe and Zn contents suggests good prospects of simultaneous improvement for both micronutrients. Among the 210 RILs, RIL 69, RIL 186, RIL 191, RIL 149 and RIL 45 were found to be more stable with higher mean micronutrient content, additive main effects and multiplicative interaction stability value (ASV) and genotype selection index (GSI) under rainfed condition. These RILs are promising and can be tested further for their combining ability for yield as well as grain micronutrient content for developing superior biofortified, heterotic pearl millet hybrids.